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Enzymatic Catalysis (enzymatic + catalysis)

Distribution by Scientific Domains

Chemistry	63%
Life Sciences	27%

Selected Abstracts

Effect of Modified Enzymatic Catalysis on the Extraction of Diosgenin from Dioscorea zingiberensis C.,H.

CHEMICAL ENGINEERING & TECHNOLOGY (CET), Issue 11 2007
Wright
Abstract Multi-enzymatic catalysis combined with acid hydrolysis is studied in order to enhance the efficiency of the enzymatic catalysis and reduce the mass transfer resistance from starch and cellulose in the extraction of diosgenin from Dioscorea zingiberensis C.,H. Wright. The cellulase is modified by polyethylene to increase its optimal reaction temperature and pH value. The modified cellulase shows better thermostability and resistance to alkali. The modified cellulase, , -amylase and , -glycosidase are used to construct the multi-enzyme and multi-enzyme catalysis is used as a pretreatment process. Compared to primary industrial techniques including acid hydrolysis, spontaneous fermentation and enzymatic catalysis, conventional techniques are optimized by using multi-enzymatic catalysis together with acid hydrolysis because of the higher reaction efficiency and lower levels of manipulation required. The purity of the product is more than 96,% with this technique, and the melting point is 205,207,°C. The diosgenin yield rate and the extraction rate reached are 2.43,% and 98,%, respectively. IR and 1H NMR spectroscopy were used to confirm the structure of the product. [source]

Optimization of Reaction Conditions for Enzymatic Viscosity Reduction and Hydrolysis of Wheat Arabinoxylan in an Industrial Ethanol Fermentation Residue

BIOTECHNOLOGY PROGRESS, Issue 2 2006
Hanne R. Sørensen
This study examined enzyme-catalyzed viscosity reduction and evaluated the effects of substrate dry matter concentration on enzymatic degradation of arabinoxylan in a fermentation residue, "vinasse", resulting from industrial ethanol manufacture on wheat. Enzymatic catalysis was accomplished with a 50:50 mixture of an enzyme preparation from Humicola insolens, Ultraflo L, and a cellulolytic enzyme preparation from Trichoderma reesei, Celluclast 1.5 L. This enzyme mixture was previously shown to exhibit a synergistic action on arabinoxylan degradation. The viscosity of vinasse decreased with increased enzyme dosage and treatment time at pH 5, 50 °C, 5 wt % vinasse dry matter. After 24 h of enzymatic treatment, 76,84%, 75,80%, and 43,47%, respectively, of the theoretically maximal arabinose, xylose, and glucose releases were achieved, indicating that the viscosity decrease was a result of enzyme-catalyzed hydrolysis of arabinoxylan, ,-glucan, and cellulose. In designed response surface experiments, the optimal enzyme reaction conditions with respect to pH and temperature of the vinasse, the vinasse supernatant (mainly soluble material), and the vinasse sediment (mainly insoluble substances) varied from pH 5.2,6.4 and 41,49 °C for arabinose release and from pH 4.9,5.3 and 42,46 °C for xylose release. Even though only limited hydrolysis of the arabinoxylan in the vinasse sediment fraction was obtained, the results indicated that the same enzyme activities acted on the arabinoxylan in the different vinasse fractions irrespective of the state of solubility of the substrate material. The levels of liberated arabinose and xylose increased with increased dry matter concentration during enzymatic hydrolysis in the vinasse and the vinasse supernatant, but at the same time, increased substrate dry matter concentrations gave corresponding linear decreases in the hydrolytic efficiency as evaluated from levels of monosaccharide release per weight unit dry matter. The study thus documents that enzymatic arabinoxylan hydrolysis of the vinasse significantly decreases the vinasse viscosity and that a compromise in the dry matter must be found if enzymatic efficiency must be balanced with monosaccharide yields. [source]

Application of Exchangeable Biochemical Reactors with Oxidase-Catalase-Co-immobilizates and Immobilized Microorganisms in a Microfluidic Chip-Calorimeter

ENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 5 2008
M. Leifheit
Abstract Several methods for the quantitative detection of different compounds, e.g., L -amino acids, sugars or alcohols in liquid media were developed by application of an automatic measuring unit including a fluid chip-calorimeter FCC-21. For this purpose, enzymes were immobilized covalently on the inner and outer surface of CPG (controlled porous glass)-spherules with an outer diameter of 100,,m and filled into a micro flow-through reaction chamber (VR = 20,,L). The design of the measuring cell allows for easy insertion into the calorimeter device of a stored series of comfortably pre-fabricated measuring cells. These cells can be filled with different enzyme immobilizates. Different oxidases were used and co-immobilized with catalase for the improvement of the detection sensitivity. A signal amplification could be achieved up to a factor of 3.5 with this configuration. ,- D -glucose, ethanol and L -lysine could be detected in a range of 0.25,1.75,mM using glucose oxidase, alcohol oxidase and lysine oxidase. The group of oxidases in combination with the enzymatic catalysis of the intermediate H2O2 allows the quantitative detection of a large number of analytes. A good measurement and storage stability could be achieved for several weeks by this immobilization method. In addition to enzyme-based detection reactions, it was shown that living microorganisms can be immobilized in the reaction chamber. Thus, the system can be used as a whole-cell biosensor. The quantitative detection of phenol in the range of 10,100,,M could be performed using the actinomycete Rhodococcus sp. immobilized on glass beads by means of embedding into polymers. [source]

SN2 Displacement by Bromide Ions in Dichloromethane , The Role of Reverse Micelles

EUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 18 2006
Lucia Brinchi
Abstract Reverse micellar systems are of interest as reaction media because they are powerful models for biological compartmentalization, enzymatic catalysis and separation of biomolecules. Solutions of ionic surfactants in apolar solvents may contain reverse micelles, but they may also contain ion pairs, or small clusters, with waters of hydration. We studied the bimolecular reaction in CH2Cl2 solutions of cationic tetraalkylammonium bromide salts (onium salts), such as cetyltrimethylammonium bromide (CTABr), cetyltripropylammonium bromide (CTPABr) and tetra- n -butylammonium bromide (TBABr). Methylnaphthalene-2-sulfonate (,-MeONs), its 6-sulfonate derivative (,-MeONsS,) as the 2,6-lutidinium salt and methyl-5- N,N,N,trimethylammonium naphthalene-1-sulfonate (,-MeONsNT+) as the trifluoromethanesulfonate salt react with Br, in CH2Cl2. First-order rate constants, kobs, increase linearly and similarly for the three substrates with increasing concentrations of the onium salts. Reactions are faster with TBABr than they are with CTPABr and CTABr, and the reactivity of the three substrates is in the order: ,-MeONsNT+ >> ,-MeONsS, > ,-MeONs. The reactions are inhibited by the addition of H2O, but CTABr tolerates H2O in large excess. At [H2O]/[CTABr] = w0 , 6, "water-pool" reverse micelles form, and kobs for all three substrates is then independent of w0. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2006) [source]

Functional characterization of an orphan cupin protein from Burkholderia xenovorans reveals a mononuclear nonheme Fe2+ -dependent oxygenase that cleaves ,-diketones

FEBS JOURNAL, Issue 20 2009
Stefan Leitgeb
Cupins constitute a large and widespread superfamily of ,-barrel proteins in which a mononuclear metal site is both a conserved feature of the structure and a source of functional diversity. Metal-binding residues are contributed from two core motifs that provide the signature for the superfamily. On the basis of conservation of this two-motif structure, we have identified an ORF in the genome of Burkholderia xenovorans that encodes a novel cupin protein (Bxe_A2876) of unknown function. Recombinant Bxe_A2876, as isolated from Escherichia coli cell extract, was a homotetramer in solution, and showed mixed fractional occupancy of its 16.1 kDa subunit with metal ligands (0.06 copper; 0.11 iron; 0.17 zinc). Our quest for possible catalytic functions of Bxe_A2876 focused on Cu2+ and Fe2+ oxygenase activities known from related cupin enzymes. Fe2+ elicited enzymatic catalysis of O2 -dependent conversion of various ,-diketone substrates via a nucleophilic mechanism of carbon,carbon bond cleavage. Data from X-ray absorption spectroscopy (XAS) support a five-coordinate or six-coordinate Fe2+ center where the metal is bound by three imidazole nitrogen atoms at 1.98 Å. Results of structure modeling studies suggest that His60, His62 and His102 are the coordinating residues. In the ,best-fit' model, one or two oxygens from water and a carboxylate oxygen (presumably from Glu96) are further ligands of Fe2+ at estimated distances of 2.04 Å and 2.08 Å, respectively. The three-histidine Fe2+ site of Bxe_A2876 is compared to the mononuclear nonheme Fe2+ centers of the structurally related cysteine dioxygenase and acireductone dioxygenase, which also use a facial triad of histidines for binding of their metal cofactor but promote entirely different substrate transformations. [source]

The phosphate site of trehalose phosphorylase from Schizophyllum commune probed by site-directed mutagenesis and chemical rescue studies

FEBS JOURNAL, Issue 5 2008
Christiane Goedl
Schizophyllum commune,,,-trehalose phosphorylase utilizes a glycosyltransferase-like catalytic mechanism to convert its disaccharide substrate into ,- d -glucose 1-phosphate and ,- d -glucose. Recruitment of phosphate by the free enzyme induces ,,,-trehalose binding recognition and promotes the catalytic steps. Like the structurally related glycogen phosphorylase and other retaining glycosyltransferases of fold family GT-B, the trehalose phosphorylase contains an Arg507-XXXX-Lys512 consensus motif (where X is any amino acid) comprising key residues of its putative phosphate-binding sub-site. Loss of wild-type catalytic efficiency for reaction with phosphate (kcat/Km = 21 000 m,1·s,1) was dramatic (,107 -fold) in purified Arg507,Ala (R507A) and Lys512,Ala (K512A) enzymes, reflecting a corresponding change of comparable magnitude in kcat (Arg507) and Km (Lys512). External amine and guanidine derivatives selectively enhanced the activity of the K512A mutant and the R507A mutant respectively. Analysis of the pH dependence of chemical rescue of the K512A mutant by propargylamine suggested that unprotonated amine in combination with H2PO4,, the protonic form of phosphate presumably utilized in enzymatic catalysis, caused restoration of activity. Transition state-like inhibition of the wild-type enzyme A by vanadate in combination with ,,,-trehalose (Ki = 0.4 ,m) was completely disrupted in the R507A mutant but only weakened in the K512A mutant (Ki = 300 ,m). Phosphate (50 mm) enhanced the basal hydrolase activity of the K512A mutant toward ,,,-trehalose by 60% but caused its total suppression in wild-type and R507A enzymes. The results portray differential roles for the side chains of Lys512 and Arg507 in trehalose phosphorylase catalysis, reactant state binding of phosphate and selective stabilization of the transition state respectively. [source]

Synthesis and characterization of metal binding pseudotripeptides

JOURNAL OF PEPTIDE SCIENCE, Issue 8 2003
Sebastian Kuenzel
Abstract Metal complexes with peptide or pseudopeptide type ligands can serve as good model compounds for a deeper understanding of enzymatic catalysis, but ligands with a high selectivity for different transition metal cations are hard to find due to the rather flexible nature of peptides. Since such ligands would be the sine qua non condition for the synthesis of heterodinuclear peptide metal complexes with catalytic activity, the search for small, affine and selective metal chelating sequences is of interest. Using four different amino acids (His, Lys, Asp, Glu) a set of 16 pseudotripeptides of the common structure Bz-AS1 -Sar-AS2 -NH2 has been synthesized, purified and characterized by mass spectrometry and 1H-NMR. Their ability to form metal complexes has been investigated leading to short motifs capable of selectively binding only one or two transition metal cations with high affinity. As expected, the complexation of transition metal cations by pseudotripeptides is strongly dependent not only on the amino acid composition, but also on the sequence with regard to the stability of the resulting complexes, as well as the selectivity of the ligands towards Cu2+, Co2+, Ni2+, Zn2+ and Mn2+. Copyright © 2003 European Peptide Society and John Wiley & Sons, Ltd. [source]

Green polymer chemistry using nature's catalysts, enzymes

JOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 12 2009
Judit E. Puskas
Abstract The use of enzymes as catalysts for organic synthesis has become an increasingly attractive alternative to conventional chemical catalysis. Enzymes offer several advantages including high selectivity, ability to operate under mild conditions, catalyst recyclability, and biocompatibility. Although there are many examples in the literature involving enzymes for the synthesis of polymers, our search showed that very little had been done in the area of polymer modification. In this article, we will discuss enzyme catalysis in general and highlight our recent results concerning precision polymer functionalization using enzymatic catalysis,"green polymer chemistry." © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47: 2959,2976, 2009 [source]

Edited by K. Morokuma and D. Musaev Computational modeling for homogeneous and enzymatic catalysis: a knowledge-base for designing efficient catalysts Wiley,VCH, 2008, 398 pp. (hardover) ISBN 978-3-527-31843-8

APPLIED ORGANOMETALLIC CHEMISTRY, Issue 11 2009
Hans Martin Senn
No abstract is available for this article. [source]

Exploring multiplicity conditions in enzymatic reaction networks

BIOTECHNOLOGY PROGRESS, Issue 3 2009
Irene Otero-Muras
Abstract In this work, a novel algorithmic approach to detect multiplicity of steady states in enzymatic reaction networks is presented. The method exploits the structural properties of networks derived from the Chemical Reaction Network Theory. In first instance, the space of parameters is divided in different regions according to the qualitative behavior induced by the parameters in the long term dynamics of the network. Once the regions are identified, a condition for the appearance of multiplicities is checked in the different regions by solving a given optimization problem. In this way, the method allows the characterization of the whole parameter space of biochemical networks in terms of the appearance or not of multistability. The approach is illustrated through a well-known case of enzymatic catalysis with substrate inhibition. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 [source]